PU.1对牙龈卟啉单胞菌脂多糖刺激下衰老巨噬细胞产生凋亡抵抗的调控作用  

Regulation of PU.1 on apoptosis resistance of aging macrophages stimulated by Porphyromonas gingivalis lipopolysaccharide

作　　者：方慧 袁怡婷 张永春 任山杉 陈露露 廖维 田艾 FANG Hui;YUAN Yiting;ZHANG Yongchun;REN Shanshan;CHEN Lulu;LIAO Wei;TIAN Ai(School of Stomatology,Guizhou Medical University,Guiyang 550004,Guizhou,China;不详)

机构地区：[1]贵州医科大学口腔医学院,贵州贵阳550004 [2]贵州医科大学附属口腔医院口腔修复种植科,贵州贵阳550004

出　　处：《实用医学杂志》2025年第4期471-477,共7页The Journal of Practical Medicine

基　　金：国家自然科学基金项目(编号:82260193);贵州省科学技术基金资助项目(编号:黔科合基础-ZK[2024]一般251);贵州省卫生健康委科学技术基金资助项目(编号:gzwkj2024-193)。

摘　　要：目的探讨在牙龈卟啉单胞菌(Porphyromonas gingivalis)脂多糖(LPS)(P.g⁃LPS)模拟的体外炎性刺激下,PU.1对衰老巨噬细胞产生凋亡抵抗的调控作用。方法利用GEO数据库分析PU.1在牙周炎牙龈组织与正常牙龈组织中的表达;将小鼠巨噬细胞系(RAW264.7)设为control组、P.g-LPS组、H_(2)O_(2)组、PU.1抑制剂组,采用RT⁃qPCR检测衰老相关分泌表型(IL-6、IL-1β、TNF-α)以及PU.1的mRNA表达;蛋白质印迹法检测p21、p16、BAX、caspase-3、Bcl-2、Bcl-xl以及PU.1蛋白表达;衰老相关β-半乳糖苷酶(senes⁃cence⁃associated⁃β⁃galactosidase,SA⁃β⁃gal)染色检测衰老细胞数量;流式细胞仪检测细胞凋亡率。结果与control组相比,P.g-LPS组和H_(2)O_(2)组p21、p16蛋白表达上调,IL-6、IL-1β、TNF-α的mRNA上调,衰老细胞数量增加,Bcl-2、Bcl-xl表达上调、BAX、caspase-3表达减少,细胞凋亡率减少,同时伴随PU.1 mRNA以及蛋白表达升高(P<0.05);与P.g-LPS组相比,PU.1抑制剂组PU.1的mRNA及蛋白表达均下调,且Bcl-2、Bcl-xl表达下调,BAX、caspase-3表达增加,细胞凋亡率升高,同时衰老细胞数量减少,IL-6、IL-1β、TNF-α的mRNA降低,p21、p16蛋白表达下调(P<0.05)。结论体外炎性刺激下,上调PU.1诱导衰老巨噬细胞产生凋亡抵抗;抑制PU.1可促进衰老巨噬细胞凋亡,减少衰老巨噬细胞数量,并下调炎性因子分泌。Objective To investigate the regulatory effect of PU.1 on apoptosis resistance of aging macro⁃phages under in vitro inflammatory stimulation simulated by lipopolysaccharide(LPS)of Porphyromonas.Methods The expression of PU.1 in periodontitis gingival tissue and normal gingival tissue was analyzed by GEO database.Mouse macrophage cell line RAW264.7 was divided into control group,P.g-LPS group,H_(2)O_(2) group and PU.1 inhibitor group.mRNA expression of senescence associated secretory phenotype(IL-6,IL-1β,TNF-α)and PU.1 were detected by RT⁃qPCR;The protein expressions of p21,p16,BAX,caspase-3,Bcl-2,Bcl-xl and PU.1 were detected by Western blot.The number of senescent cells was detected by senescence⁃associated⁃galactosidase(SA⁃β⁃gal)staining.The apoptosis rate was detected by flow cytometry.Results Compared with control group,the expression of p21,p16 protein and mRNA of IL-6,IL-1βand TNF-αwere up-regulated in P.g-LPS group and H_(2)O_(2) group,the number of senescent cells was increased,the expression of Bcl-2 and Bcl-xl was up-regulated,the expression of BAX and caspase-3 was decreased,and the apoptosis rate was decreased.Meanwhile,the mRNA and protein expression of PU.1 were increased(P<0.05).Compared with P.g-LPS group,mRNA and protein expression of PU.1 in PU.1 inhibitor group were down-regulated,Bcl-2 and Bcl-xl were down-regulated,BAX and caspase-3 expressions were increased,apoptosis rate was increased,the number of senescent cells was decreased,and mRNA levels of IL-6,IL-1βand TNF-αwere decreased.The expression of p21 and p16 proteins were downregulated.(P<0.05).Conclusion Under inflammatory stimulation in vitro,increased expression of PU.1 induced apoptosis resistance of aging macrophages,inhibition of PU.1 promoted apoptosis of aging macrophages,reduced the number of aging macrophages,and down-regulated the secretion of inflammatory factors.

关 键 词：PU.1 巨噬细胞 细胞衰老 P.g-LPS 

分 类 号：R781.42[医药卫生—口腔医学]